Methods for the identification of inhibitors of α-aminoadipate reductase as antibiotics

A method for identifying a test compound as a candidate for an antibiotic, comprising:

• a) contacting L-2-Aminoadipate and NADPH and ATP with an α-Aminoadipate Reductase;
• b) contacting L-2-Aminoadipate and NADPH and ATP with α-Aminoadipate Reductase and said test compound;
• and
• c) determining the change in concentration for at least one of the following:
• L-2-Aminoadipate, L-2-Aminoadipate 6-semialdehyde, NADP+, NADPH, AMP, ATP, H2O and/or pyrophosphate;
• wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.

A method for identifying a test compound as a candidate for an antibiotic, comprising:

• a) contacting L-2-Aminoadipate 6-semialdehyde, NADP+, AMP, pyrophosphate, and H2O with an α-Aminoadipate Reductase;
• b) contacting L-2-Aminoadipate 6-semialdehyde, NADP+, AMP, pyrophosphate, and H2O with an α-Aminoadipate Reductase and said test compound;
• and
• c) determining the change in concentration for at least one of the following:
• L-2-Aminoadipate, L-2-Aminoadipate 6-semialdehyde, NADP+, NADPH, AMP, ATP, H2O and/or pyrophosphate;
• wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.

A method for identifying a test compound as a candidate for an antibiotic, comprising:

• a) contacting L-2-Aminoadipate and NADPH and ATP with a polypeptide selected from the group consisting of:
• a polypeptide having at least 50% sequence identity with an α-Aminoadipate Reductase, a polypeptide having at least 50% sequence identity with an α-Aminoadipate Reductase and having at least 10% of the activity thereof, and a polypeptide comprising at least 100 consecutive amino acids of an α-Aminoadipate Reductase
• b) contacting L-2-Aminoadipate and NADPH and ATP with said polypeptide and said test compound;
• and
• c) determining the change in concentration for at least one of the following:
• L-2-Aminoadipate, L-2-Aminoadipate 6-semialdehyde, NADP+, NADPH, AMP, ATP, H2O and/or pyrophosphate;
• wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.

A method for identifying a test compound as a candidate for an antibiotic, comprising:

• a) contacting L-2-Aminoadipate 6-semialdehyde, NADP+, AMP, pyrophosphate, and H2O with a polypeptide selected from the group consisting of:
• a polypeptide having at least 50% sequence identity with an α-Aminoadipate Reductase, a polypeptide having at least 50% sequence identity with an α-Aminoadipate Reductase and at least 10% of the activity thereof, and a polypeptide comprising at least 100 consecutive amino acids of an α-Aminoadipate Reductase
• b) contacting L-2-Aminoadipate 6-semialdehyde, NADP+, AMP, pyrophosphate, and H2O with said polypeptide and said test compound;
• and
• c) determining the change in concentration for at least one of the following:
• L-2-Aminoadipate, L-2-Aminoadipate 6-semialdehyde, NADP+, NADPH, AMP, ATP, H2O and/or pyrophosphate;
• wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.

A method for identifying a test compound as a candidate for an antibiotic, comprising:

• a) measuring the expression of a Magnaporthe α-Aminoadipate Reductase in a cell, cells, tissue, or an organism in the absence of said compound;
• b) contacting said cell, cells, tissue, or organism with said test compound and measuring the expression of said Magnaporthe α-Aminoadipate Reductase in said fungus or fungal cell;
• c) comparing the expression of Magnaporthe α-Aminoadipate Reductase in steps (a) and (b);
• wherein a lower expression in the presence of said test compound indicates that said compound is a candidate for an antibiotic.

A method for identifying a test compound as a candidate for an antibiotic, comprising:

• a) providing cells having one form of a gene in the lysine biochemical and/or genetic pathway and providing comparison cells having a different form of said gene,
• b) contacting said cells and comparison cells with a said test compound,
• c) determining the growth of said cells and comparison cells in the presence of said test compound;
• wherein a difference in growth between said cells and said comparison cells in the presence of said compound indicates that said compound is a candidate for an antibiotic.

A method for identifying a test compound as a candidate for an antibiotic, comprising:

• (a) providing paired growth media;
• comprising a first medium and a second medium, wherein said second medium contains a higher level of lysine than said first medium;
• (b) contacting an organism with said test compound;
• (c) inoculating said first and second media with said organism;
• and
• (d) determining the growth of said organism;
• wherein a difference in growth of the organism between said first and second media indicates that said test compound is a candidate for an antibiotic.